摘要
目的探索一种改进的贴壁培养神经干细胞的方法。方法从胚胎期为15.5d的胎鼠大脑皮层中提取神经干细胞,在PO/FN(Poly-L-ornithine hydrochloride/Fibronectin)包被处理后的培养瓶中进行贴壁培养并传代,在倒置显微镜下观察神经干细胞的细胞形态,并通过免疫荧光法鉴定神经干细胞及其子代细胞进行分化操作之后的情况。结果原代神经干细胞细胞形态清晰,免疫荧光标记物明显。结论相对于传统神经球悬浮培养法该法处理简便,传代损耗低,容易观察到细胞形态和分化阶段,更有助于神经干细胞的培养和存储。
Objective To develop a modified method of adherent culturing NSCs to replace the traditional suspension culturing method.Methods Neural stem cells（NSCs）were isolated from brain of fetal mouse at 15.5-day fetal age.In culture bottle which had been pre-coated with poly-L-ornithine hydrochloride/fibronectin（PO/FN）,the cells were adherently cultured and passaged.The cell morphology was observed under inverted microscope.NSCs and their differentiated cells were identified by immunofluorescence.Results NSCs obtained in this study were successfully adherently cultured and expressed specific markers.Conclusion NSCs are successfully adherently cultured in this study.As compared with the traditional suspension culturing method,adherent culturing is simpler and has lower wastage of passage.It is easy to observe cell morphology and differentiation by this method.It is also helpful to culturing and storage of NSCs.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2017年第3期312-316,340,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81270700)
关键词
神经干细胞
贴壁培养
包被处理
免疫荧光
悬浮培养
neural stem cells
adherent culture
coating process
immunofluorescence
suspension culture
